LOCUS       sequence-178838-        4652 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pin nuclease domain with Pum cloning site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4652)
  AUTHORS   Adamala KP, Martin-Alarcon DA, Boyden ES
  TITLE     Programmable RNA-binding protein composed of repeats of a single 
            modular unit.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 May 10;113(19):E2579-88. doi: 
            10.1073/pnas.1519368113. Epub 2016 Apr 26.
  PUBMED    27118836
REFERENCE   2  (bases 1 to 4652)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1073/pnas.1519368113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-05-10- 10"; volume: "113"; issue: "19"; pages: 
            "E2579-88"
COMMENT     Sequence Label: [GG][Pin]
FEATURES             Location/Qualifiers
     source          1..4652
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..29
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    743..864
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(754..773)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     808..827
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      1045..1500
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1132..1151)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1342..1363
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1567..1586)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1686..1708
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1746..1764)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1832..1936
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1937..2005,2006..2797)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2155..2174)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2968..3556
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3457..3476
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3767..4146
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4147..4350
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4301..4321
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4347..4371
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          4486..4618
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4601..4620
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 ctagagtact taatacgact cactataggc tagcctcgag cgccaccatg cgagagagac
       61 cggatggcag agagtcgcta tctcatgtac cgatagtagc ggtctccacg cgagtggaca
      121 ccggaaacgg ctccatggaa ctcgagatca ggccactttt cctggtgcca gacactaatg
      181 gattcataga ccacctcgcc agcctcgcac gcctcttgga gtcccgcaag tatattctgg
      241 tagttccgct gatcgttatc aatgagttgg acggcttggc caagggacag gagactgacc
      301 atcgcgccgg tgggtacgcc cgcgtcgtgc aagagaaggc tagaaaatca atcgaattcc
      361 tggagcaacg cttcgaatca cgggattcct gcctgagggc tcttacctcc agaggcaacg
      421 agctggagtc tattgcattc cgcagcgagg acatcacagg gcagctgggc aataacgatg
      481 atcttattct gagctgttgt cttcactatt gtaaggacaa ggctaaagat ttcatgcccg
      541 ctagtaagga agaacccatt cggctgctgc gcgaagtcgt gctgctgaca gacgatcgca
      601 atctccgggt taaagccttg actcggaacg tgccagtgag ggatatcccc gccttcctga
      661 catgggctca ggtgggataa gaattcgtaa cacgcgtggt acctctagag tcgacccggg
      721 cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga
      781 atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc
      841 attataagct gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt
      901 cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc
      961 gataaggatc cgggctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     1021 ttgcgcagcc tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     1081 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     1141 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     1201 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     1261 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     1321 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     1381 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     1441 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     1501 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac
     1561 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     1621 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     1681 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     1741 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     1801 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1861 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1921 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1981 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2041 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     2101 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2161 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2221 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2281 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2341 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     2401 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     2461 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     2521 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2581 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2641 cggtgagcgt gggtcgcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2701 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2761 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2821 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2881 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2941 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3001 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3061 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     3121 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3181 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3241 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3301 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3361 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3421 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3481 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3541 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3601 gccttttgct cacatggctc gacagatctt caatattggc cattagccat attattcatt
     3661 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     3721 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttggca ttgattattg
     3781 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     3841 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     3901 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     3961 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     4021 ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     4081 tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     4141 accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg
     4201 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     4261 cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg cggtaggcgt
     4321 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat cactagaagc
     4381 tttattgcgg tagtttatca cagttaaatt gctaacgcag tcagtgcttc tgacacaaca
     4441 gtctcgaact taagctgcag aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt
     4501 acaagacagg tttaaggagg ccaatagaaa ctgggcttgt cgagacagag aagactcttg
     4561 cgtttctgat aggcacctat tggtcttact gacatccact ttgcctttct ctccacaggt
     4621 gtccactccc agttcaatta cagctcttaa gg
//
